Raman microspectroscopic study on polymerization and degradation processes of a diacetylene derivative at surface enhanced Raman scattering active substrates. 1. Reaction kinetics.

Raman microspectroscopy was applied to study the polymerization kinetics of the Langmuir-Blodgett (LB) films of 10,12-pentacosadiynoic acid (DA) adsorbed on surface enhanced Raman scattering (SERS) active Ag island films. A two-dimensional (2D) Raman microscopic image measured at 1520 cm(-1) exhibits bright and dim spots with the diameter of several hundred nanometers. Raman microscopic spectra, measured by defocusing the excitation laser light (532 nm, diameter of ca. 10 mum) on the samples at room temperature, proved the occurrence of the surface processes consisting of the formation of polydiacetylene (PDA) in the blue phase, its conversion to the red phase, and subsequent bleaching. These reactions were negligible under the same condition for the DA-LB films prepared on a smooth (i.e., SERS-inactive) Ag film, indicating that the 532-nm-induced polymerization and the bleaching process are enhanced by the SERS-active substrates. At low temperatures below -50 degrees C, the Raman microscopic measurements proved the formation of the blue phase and its conversion to the red phase with much lower reaction rates compared to the corresponding rates at room temperature. The bleaching, however, was much suppressed at the low temperatures. The kinetic analyses of the formation of the blue phase and its conversion to the red phase were performed by using the intensity changes of the Raman bands due to the blue (1477 cm(-1)) and red (1517 cm(-1)) phases as a function of the irradiation time. The results strongly suggested the existence of at least two processes taking place simultaneously on the SERS-active substrates; that is, one of the processes is a sequential reaction, DA-monomers --> PDA in the blue phase --> PDA in the red phase, and the other is another sequential reaction, DA-monomers --> PDA in the red phase --> degradation species (probably amorphous carbon). Thus, even at the low temperatures, there occurs the surface reaction consisting of the formation of PDA and its degradation. The reaction can be ascribed to a process taking place at the highly SERS-active site, which gives the bright spot (so-called "hot spot") on the 2D Raman image, as proved by the confocal Raman microscopic measurement in the following paper.

Raman microspectroscopic study on polymerization and degradation processes of a diacetylene derivative at surface enhanced Raman scattering active substrates. 2. Confocal Raman microscopic observation of polydiacetylene adsorbed on active sites.

Confocal Raman microscopic measurements were performed at room temperature on the Langmuir-Blodgett (LB) monolayer of 10,12-pentacosadiynoic acid (DA) prepared on surface enhanced Raman scattering (SERS) active Ag island films, two-dimensional (2D) Raman images of which exhibit bright and dim spots on a dark background. The measurements performed by focusing the excitation laser light (488 nm) on the dark background indicate the prompt appearance of the Raman bands (1515 and 2115 cm(-1)) due to polydiacetylene (PDA) in the red phase and subsequent diminution of the Raman bands. On the other hand, the spectra observed by focusing the excitation laser spot on the dim and bright spots exhibit almost random fluctuations, giving rather narrow Raman bands in the 1620-1000 cm(-1) region, which appear and disappear temporarily with varying intensities under the continuous irradiation at 488 nm. Broad Raman bands appear around 1580 and 1360 cm(-1), which are ascribable to amorphous carbon, at a later stage of the observation, the intensities from the bright spot being more than 100 times stronger than those from the dim spot. The narrow bands are ascribed to a series of carbonaceous intermediates such as polyenes, graphite sheets with various sizes, and folded or reorganized forms of the sheets including carbon nanotubes and fullerenes, which are formed during the conversion of PDA to amorphous carbon. The random spectral fluctuation was interpreted by considering that the intermediates undergo thermally activated diffusion and get temporarily in contact with the SERS-active site, resulting in the enhancement of their Raman bands and the fluctuation.

Metastatic patterns of prostatic cancer. Correlation between sites and number of organs involved.

Prostatic cancer in 1885 autopsy cases was classified according to the number of organs involved in metastasis, and a comparison was made concerning the frequency of metastasis to the various organs. The frequencies of metastasis to the lungs and para-aortic lymph nodes were low in cases with single-organ involvement (4.6% and 2.3%, respectively), but increased rectilinearly in accordance with the number of organs involved and became high in cases with metastasis to three or more organs (49.1% and 21.8% in total, respectively). On the other hand, the frequencies of local extension to the bladder and invasion of the pelvic lymph nodes were high even in cases with single-organ involvement (34.5% and 9.2%, respectively) and were not significantly changed regardless of the number of organs involved. No significant correlation was seen between pelvic and para-aortic lymph node involvement. In cases with single-organ involvement, metastasis to the lumbar spine occurred frequently, but those to the ribs, sternum, and ilium occurred less frequently. There may be multiple metastases in cases with metastases to the para-aortic lymph nodes, sternum, and ilium. The number of metastatically involved organs is useful in analyzing the mode of metastasis.

[Combination of intra-arterial administration of anticancer agent and hemocarboperfusion in treatment of advanced cancer].

Nineteen patients with malignant tumors were treated with a combination of bolus administration of mitomycin C with direct hemoperfusion over charcoal, and 15 patients were valuable. Mitomycin C of 1 mg/kg was administered via a catheter placed into the regional artery. Seven out of 14 patients with genitourinary cancers: 4 out of 9 patients with urinary bladder, 2 of 4 patients with renal and one prostate responded to the therapy. There were 4 responders out of 8 in the one-shot administration group, on the other hand, 7 cases receiving continuous infusion of 60 to 120 minutes responded. In both the one-shot group and the continuous group nausea and vomiting were observed in 21% and 11% respectively. Three patients receiving continuous administration of mitomycin C had local ulcerations. Myelosuppressions such as leukocytopenia and thrombocytopenia were observed in 37% of the both groups.

The relationship of renal histopathological lesions to immunoglobulin classes and complement fixation of anti-native DNA antibodies in systemic lupus erythematosus.

The relationship of renal immunohistopathological lesions to the immunoglobulin classes (Ig) and complement fixation (CF) of anti-native DNA antibodies (anti-nDNA) in sea was studied in 20 patients with systemic lupus erythematosus (SLE) on whom renal biopsy was performed. The Ig classes and CF of anti-nDNA were assessed by the indirect immunofluorescent and complement fixation methods employing kinetoplasts of Crithidia luciliae as substrate. The histopathologically active lesions of lupus nephritis showed a significant correlation with the IgG and complement fixation of anti-native DNA antibodies in sera. IgM anti-nDNA superiority was not necessarily observed in patients with inactive histological lesions, but rather in those with thickening of the glomerular basement membrane. The granular deposits of immunoglobulin or complement in the renal glomeruli showed a significant correlation with the presence of IgG and CF of anti-nDNA in sera, while the deposition of C3 in the glomeruli also showed a significant correlation with the CF of anti-nDNA.

Avidity indices of anti-IgG antibodies in diseases.

Anti-IgG antibodies were detected in 69% of 235 sera tested. A simple and reproducible method was developed to measure the avidity index of anti-IgG antibodies. It was found that subjects with systemic lupus erythematosus and progressive systemic sclerosis and normal controls had antibodies with lower avidity index than those with rheumatoid arthritis and Down's syndrome. The difference in avidity seems to be disease specific. Avidity does not correlate with antibody titers, but all sera with high avidity antibody are positive in the latex test. The anti-IgG as measured by this method was shown to be mainly of IgM class. The avidity indices of whole sera and purified IgM fractions were similar.

Superoxide assay-leukocyte adherence inhibition test and a soluble factor which stimulates the adherence of macrophages.

A modified method for the leukocyte adherence inhibition test is described. Peritoneal cells from immune guinea pigs or peripheral mononuclear cells from cancer patients were incubated with immunizing antigen or tumor extract in a 4-mm-wide glass microcell for spectrophotometer analysis. Instead of visual cell counting, the cells adherent to the bottom of the microcell were stimulated with cytochalasin E and wheat germ agglutinin, and the amount of the superoxide (O2.-) generated from the adherent macrophages or monocytes was assayed. Antigen-specific adherence inhibition of peritoneal cells of the immunized guinea pigs was detected 2 weeks after immunization. In contrast, cell adherence was stimulated after 3 weeks. It was shown that a soluble factor was responsible for the adherence stimulation and that nonadherent cells were necessary for its production. Peripheral mononuclear cells of 70% of the tumor-bearing lung cancer patients reacted to the lung tumor extract (9 adherence inhibitions and 7 adherence stimulations per 23 patients). Twenty-five % (3 of 12) of tumor-free patients showed positive reactions, all with adherence stimulation. Of the 12 healthy donors, 3 cases of pneumonia, one case of angiosarcoma of the left flank, one case of hemangiopericytoma of the mediastinum, and 8 cases of breast cancer, non reacted with the lung tumor extract, and only one of 7 lung tuberculosis patients showed positive adherence stimulation.

Steroid hormone receptors and clinical usefulness in human breast cancer.

For more precise prediction of response to endocrine therapy in patients with advanced breast cancer, we investigated the correlation between the presence of combination of estrogen (ER), progesterone (PgR) receptors, or combination of ER, PgR and androgen receptor (AR) and response to therapy. The patients with ER+, PgR+ tumors survived longer than those with ER+, PgR-, or ER-, PgR- tumors. A better response rate with longer survival time was obtained in patients with tumors that had three positive receptors or PgR+ and/or AR+ in addition to ER+ than in those with tumors of other combinations. According to the sequential assays of ER in tumors in several stages of breast cancer, it is likely that the positive rate of ER becomes lessened by the progression of cancer and by treatments. We stressed the significance of steroid hormone receptor assay in the mastectomy specimens for the prediction of response of therapy in case of the future recurrence of malignancy.

Plasmapheresis in treatment of rheumatic fever.

1. Three cases of rheumatic fever were treated with plasmapheresis. Improvement of abnormal clinical and laboratory findings of the patients was obtained by this procedure. 2. High levels of immune complexes, observed in the sera of patients with rheumatic fever, reduced markedly following plasmapheresis.

Adjuvant endocrine therapy for operable breast cancer using the antiestrogen mepitiostane: a preliminary report.

The steroidal antiestrogen, mepitiostane (Thioderone, 2 alpha, 3 alpha-epitio-5 alpha-androstane-17 beta-yl 1-methoxycyclopentyl ether) was administered orally to 182 patients with operable breast cancer as an adjuvant therapy after mastectomy intermittently for 3 years. The mean duration of follow-up was 32 months (6 approximately 42 months). The treatment failure ratio and the time distribution of failure among those given mepitiostane were compared with those of 96 patients treated with mitomycin C. There was a recurrence of malignancy in 11.5% (21/182) of patients treated with mepitiostane and in 15.6% (15/96) of patients given mitomycin C. The recurrence rates of the postmenopausal patients treated with mepitiostane and those treated with motomycin C were 3.8% (3/80) and 17.6% (18/102), respectively. In contrast, the malignancy relapsed in 24.0% (14/39) and in 2.4% (1/42) of the premenopausal patients given mepitiostane and those given mitomycin C, respectively. Estrogen and progestrone receptors (ER, PgR) were assayed in some of the primary breast cancers. Although there was no difference in recurrence rate between the ER-positive and -negative cases, the recurrence rate of the patients with ER(+)PgR(+) tumors treated with mepitiostane was shown to be lower than those with ER(+)PgR(-) or ER(-)PgR(-) tumors. The antiestrogen mepitiostane may thus be useful in postmenopausal patients, in whom the adjuvant chemotherapy was less effective in preventing the recurrence of malignancy.

Quantitation and characterization of soluble immune complexes precipitated from sera by polyethylene glycol (PEG).

Polyethylene glycol (PEG) was used to isolate immune complexes from sera. Complexes were then quantified and partially characterized by a variety of immunological techniques. Complexes were detected in rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), Felty's syndrome and vasculitis, but not in polymyositis, scleroderma or Behçets syndrome. The level of immune complexes correlated with disease activity in SLE and RA patients. Rheumatoid factors and anti-DNA antibodies were enriched by PEG precipitation of RA and SLE sera, respectively, thus these antibodies appeared to be present in the form of soluble immune complexes. Immune complexes usually contained all three immunoglobulin classes, IgG, IgM and IgA. Complexes from RA patients were generally larger and often contained Clq, while C3 was found almost exclusively in SLE complexes which also tended to be smaller. Such compositional differences may one day explain the differences in clinical manifestations of various immune complex-mediated diseases.

Significance of estrogen receptor assay in cytotoxic chemotherapy in relation to previous endocrine therapy of advanced breast cancer patients.

In 36 patients with advanced breast cancer, most of whom had been subjected to major endocrine ablation therapy such as bilateral adrenalectomy, correlation between the presence or absence of estrogen receptor (ER) in tumors and response to chemotherapy was investigated. Complete or partial response was obtained in 6 of 14 (42.6%) patients with ER-positive tumors and in 3 of 22 (13.6%) patients with ER-negative tumors (P = 0.1). The response to chemotherapy was not influenced by the concentration of ER in tumors. Distribution of the duration of regression readings and the survival time was not significantly different between the patients with ER-positive and -negative tumors. Some regimens which did not include adriamycin, such as MFC (mitomycin-C, 5-fluorouracil, and cytosine arabinoside) had a good effect on the ER-positive cases, whereas the response to regimens including adriamycin was not influenced by the ER positiveness of the tumors. In the classification of some clinical parameters of the patients, response of the patients with ER-negative tumors does not seem to be better than that of the patients with ER-positive tumors. Responses to the major endocrine ablation therapy and also to the subsequently carried out chemotherapy were influenced by the presence or absence of ER in tumors before the endocrine therapy.

Characterization and measurement of anti-IgG antibodies in human sera by radioimmunoassay (RIA).

A sensitive direct binding radioimmunoassay (RIA) was developed which detected low avidity anti-IgG antibodies in sera negative in the latex fixation test (LFT). IgG class antibodies could be detected and were commonly found along with IgM class antibodies. Additionally, the RIA was more reproducible than the LFT, was easily adapted to measure relative avidities of anti-IgG antibodies, and had other technical advantages over the LFT.